Figure 4.

Smad3/Smad4 C324Y oligomerization. One milligram of total lysates was obtained by MDA MB468, untransfected and transiently transfected with pEGFPC3-Smad4 wt or pEGFPC3-Smad4 C324Y, untreated or treated with 10 ng/ml TGF-β1 for 60 min. Proteins were immunoprecipitated with anti-GFP antibody, resolved under reducing conditions by 8% SDS-PAGE and subjected to immunoblot analysis using primary antibody to P-Smad3 (1:500) and GFP (1:500). Transiently transfection and equal loading were verified using primary antibodies to GFP and β-actin.